Effects of adenovirus-mediated KLF4 over-expression on osteogenic differentiation of osteoblasts <italic style="font-style: italic">in vitro</italic>

Jiajia Yang; Ping Liu; Qian Xu; Zhihua Liu; Xiaoming Li; Chao Wang; Bo Zhang; Manjing Deng

doi:10.16016/j.1000-5404.201610196

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Effects of adenovirus-mediated KLF4 over-expression on osteogenic differentiation of osteoblasts in vitro

更新时间：2026-01-09

- Effects of adenovirus-mediated KLF4 over-expression on osteogenic differentiation of osteoblasts in vitro
- Journal of Third Military Medical University Vol. 39, Issue 7, Pages: 628-634(2017)
- 作者机构：
- 作者简介：
- 基金信息：
  
  the Independent Research Project of State Key Laboratory of Trauma, Burns and Combined Injury(SKLZZ201507)
- DOI：10.16016/j.1000-5404.201610196
  CLC：
- Received：27 October 2016，
  
  Published：2017-07
- 稿件说明：
移动端阅览
Jiajia Yang, Ping Liu, Qian Xu, et al. Effects of adenovirus-mediated KLF4 over-expression on osteogenic differentiation of osteoblasts in vitro[J]. Journal of Third Military Medical University, 2017, 39(7): 628-634.
DOI：

Jiajia Yang, Ping Liu, Qian Xu, et al. Effects of adenovirus-mediated KLF4 over-expression on osteogenic differentiation of osteoblasts in vitro[J]. Journal of Third Military Medical University, 2017, 39(7): 628-634. DOI： 10.16016/j.1000-5404.201610196.

摘要

Abstract

Objective

To determine the effect of recombinant adenovirus-mediated Krüppel like factor 4 (KLF4) transfection on the osteogenic differentiation of osteoblasts

in vitro

Methods

Methods Primary mouse osteoblasts were isolated from newborn C57 mice by enzymatic digestion and then cultured and identified. Alizarin Red S staining was carried out after osteogenic induction for 21 d. The recombinant adenoviral vector carrying the KLF4 gene (Ad-KLF4) was constructed by pHBAd-EF1-MCS-GFP system. The transfection efficiency was detected by measuring the different multiplicity of infection (MOI) with flow cytometry. Osteoblasts were randomly assigned into control group (non-transfection)

GFP group (Ad-GFP transfection) and KLF4 group (Ad-KLF4 transfection). The expression of KLF4 protein was detected by Western blotting. MTT assay was used to analyze cell proliferation. Real-time PCR was used to detect the expression of osteogenic genes alkaline phosphatase (ALP)

bone sialoprotein (BSP) and runt-related transcription factor2 (RUNX2).

Results

Primary osteoblasts were triangle and polygon in shape under inverted phase contrast micr

oscope. A large number of red calcified nodules were found after Alizarin Red S staining. Restriction enzyme digestion and gene sequencing confirmed Ad-KLF4 was constructed successfully. At the optimal multiplicity of infection (MOI

100 PFU/mL)

Ad-KLF4 transfection resulted in significant increases in protein and mRNA levels of KLF4 (

0.01)

obviously increase in osteoblasts proliferation (

0.05

0.01)

and decreases in the mRNA levels of osteogenic genes ALP

BSP and RUNX2 (

0.05

0.01)

when compared with the control group and GFP group.

Conclusion

Adenovirus-mediated KLF4 over-expression can promote osteoblast proliferation and inhibit osteoblast osteogenic differentiation

and thus inhibit bone formation.

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Keywords

references

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Effects of adenovirus-mediated KLF4 over-expression on osteogenic differentiation of osteoblasts in vitro

DOI：10.16016/j.1000-5404.201610196

摘要

Abstract

关键词

Keywords

references